LTB4 synthesis inhibitors

ABSTRACT

This invention relates to a compound of the formula: ##STR1## or a pharmaceutically acceptable salt thereof wherein X is oxgen, sulfur, --CH═CH--or --CH═N--; wherein R 1  is alkyl, alkenyl or alkynyl of about 1 to 20 carbon atoms; wherein R is --CO 2  R 2 , tetrazole, methylsulfonamide or benzenesulfonamide, wherein R 2  is hydrogen, alkyl of 1 to 6 carbon atoms or a pharmaceutically acceptable cation and R 3  is hydroxyl or halogen, having utility as LTB 4  synthesis inhibitors.

This is a division of application Ser. No. 07/452,138, filed Dec. 18,1989, now U.S. Pat. No. 5,086,067.

BACKGROUND OF THE INVENTION

The present invention relates to pharmaceutical agents (compounds) whichact as leukotriene B₄ (LTB₄) synthesis inhibitors in mammals. Thecompounds inhibit LTB₄ synthesis by inhibiting phospholipase A₂ (PLA₂)activity. PLA₂ is an important enzyme in the biosynthesis ofleukotrienes as PLA₂ acts to release arachidonic acid fromphospholipids. Once released, arachidonic acid is rapidly metabolized bya variety of enzymes of the arachidonic acid cascade to produceprostaglandins, leukotrienes and related compounds. The use of thecompounds herein to inhibit PLA₂ activity thus inhibits the release ofarachidonic acid from phospholipids. The inhibition of release ofarachidonic acid similarly diminishes subsequent products in thearachidonic acid cascade, such as prostaglandins leukotrienes, andrelated compounds, including LTB₄.

LTB₄ (Formula I) is an arachidonic acid metabolite which is produced bythe 5-lipoxyqenase pathway. Pharmacologically, LTB₄ is an importantmediator of ##STR2## inflammation. LTB₄ is known to induce chemotaxis,chemokinesis, aggregation, and degranulation of leukocytes in vitro, andto induce accumulation of polymorphonuclear leukocytes, and increasevascular permeability and edema formation in vivo. Particularly highlevels of LTB₄ are detected in lesions in inflammatory diseases such asrheumatoid or spondylarthritis, gout, psoriasis, ulcerative colitis,Crohn's disease, multiple sclerosis and some respiratory diseases. Sincethe compounds herein inhibit PLA₂ and thereby LTB₄, the compounds of thepresent invention are useful in treating inflammatory conditions inmammals such as psoriasis, Crohn's disease, ulcerative colitis, multiplesclerosis and the like.

Accordingly, it is an object of this invention to produce compounds foruse as pharmaceutical agents which will exhibit LTB₄ inhibitory activityin mammals.

The pharmacology of the biologically active leukotrienes is generallydiscussed in J. Clin. Invest. 73, 889-897 (1984).

SUMMARY OF THE INVENTION

This invention relates to a compound of the formula: ##STR3## or apharmaceutically acceptable salt thereof

wherein X is oxygen, sulfur, --CH═CH--, or CH═N;

wherein Y, when present, is hydrogen or halogen wherein R¹ is alkyl,alkenyl, or alkynyl of about 1 to 20 carbon atoms;

wherein R is --CO₂ R², tetrazole, methylsulfonamide orbenzenesulfonamide; wherein R² is hydrogen, alkyl of 1 to 6 carbons or apharmaceutically acceptable cation; and

wherein R³ is hydroxyl or halogen.

This invention, more specifically, relates to a compound of the formula:##STR4## or a pharmaceutically acceptable salt thereof

wherein X is oxygen, sulfur, --CH═CH--, or CH═N; wherein Y is hydrogenor halogen wherein R¹ is alkyl, alkenyl, or alkynyl of about 1 to 20carbon atoms;

wherein R is --CO₂ R², tetrazole, methylsulfonamide, orbenzenesulfonamide;

wherein R² is hydrogen, alkyl of 1 to 6 carbons or a pharmaceuticallyacceptable cation; and

wherein R³ is hydroxyl or halogen.

This invention also more specifically, relates to a compound of theformula: ##STR5## or a pharmaceutically acceptable salt thereof

wherein X is oxygen, sulfur, --CH═CH--, or CH═N;

wherein Y is hydrogen or halogen wherein R¹ is alkyl, alkenyl, oralkynyl of about 1 to 20 carbon atoms;

wherein R is --CO₂ R², tetrazole, methylsulfonamide, orbenzenesulfonamide;

wherein R² is hydrogen, alkyl of 1 to 6 carbons or a pharmaceuticallyacceptable cation; and

wherein R³ is hydroxyl or halogen.

DETAILED DESCRIPTION

This invention encompasses compounds of Formula VI as previouslydescribed. A particularly preferred embodiment of the present inventionis encompassed by a compound of the formula: ##STR6## or apharmaceutically acceptable salt thereof,

wherein X is oxygen, sulfur, --CH═CH--, or CH═N

The term "lower alkyl" as used herein means straight or branched chainalkyls having 1-6 carbon atoms.

The term "pharmaceutically acceptable cation" as used to describe R₂refers to cations such as ammonium, sodium, potassium, lithium, calcium,magnesium, ferrous, zinc, copper, manganous, aluminum, ferric, manganic,ammonium, tetraalkyl ammonium, and the like.

The term "pharmaceutically acceptable non-toxic addition salts" referseither to those base derived salts of any compound herein having acarboxylic acid function.

The base derived salts can be derived from pharmaceutically acceptablenon toxic inorganic or organic bases. Among the inorganic bases employedto produce pharmaceutically acceptable salts are the hydroxide bases ofthe "pharmaceutically acceptable cations" disclosed above.

Among the organic bases employed to produce pharmaceutically acceptablesalts are the pharmaceutically acceptable non toxic bases of primary,secondary, and tertiary amines. Especially preferred non-toxic bases areisopropylamine, diethylamine, ethanolamine, dicyclohexylamine, choline,and caffeine.

All of the pharmaceutically acceptable non-toxic addition salts areprepared by conventional processes which are well known to those ofordinary skill in the art.

The compounds of this invention are generally prepared according to thereaction schemes I, II and III, wherein a side chain is substituted ontoa halo aromatic acid or ester moiety. By halo is meant a halogen such asbromo, iodo, fluoro or chloro. In Scheme I, the halo group isrepresented by the term "halo." By aromatic moiety is meant phenyl,thienyl or furyl, corresponding to "X" in the aryl ring being --CH═CH--,--CH═N--, --S--, and --O--.

As disclosed in Scheme I, a side chain can be added to the aromaticmoiety by performing a nucleophilic substitution of the halogen such asvia a reaction with an alkyne, CO, and Pd[O].

Hydrogenation of the triple bond, such as by introducing hydrogen overpalladium produces an enone containing side chain having the cis ortrans configuration.

The reaction Scheme III illustrates another method for the addition ofthe side chain to the aromatic moiety. Substitution of the side chainonto the monohalo-aromatic moiety to produce XV is accomplished byperforming a nucleophilic substitution at the carbon bearing the halogroup, preferably with a nucleophile, such as an alkyne, alkene, or a(tributylstannyl) alkyne, in the presence of a catalyst, such as Pd[O],in a nonpolar solvent, such as toluene, and in the presence of heat.

The biological activity possessed by the compounds of this invention wasindicated by positive results in assays for inhibition of human synovialfluid PLA₂ (HSF-PLA₂) and LTB₄ biosynthesis in HL-60 cells.

By virtue of their activity as LTB₄ synthesis inhibitors, the compoundsof Formula I are useful in treating inflammatory conditions in mammalssuch as psoriasis, Crohn's disease, ulcerative colitis, multiplesclerosis and the like. Similarly, the compounds of Formula I can beused in preventing recurring inflammatory attacks. A physician orveterinarian of ordinary skill can readily determine whether a subjectexhibits the inflammatory condition. The preferred utility relates totreatment of ulcerative colitis.

The compounds of the present invention can be administered in such oraldosage forms as tablets, capsules, softgels, pills, powders, granules,elixirs, or syrups.

The compounds can also be administered intravascularly,intraperitoneally, subcutaneously, intramuscularly, or topically usingforms known to the pharmaceutical art. Moreover, they can beadministered rectally or vaginally, in such forms as suppositories orbougies. In general, the preferred form of administration is oral. Forthe orally administered pharmaceutical compositions and methods of thepresent invention, the foregoing active ingredients will typically beadministered in admixture with suitable pharmaceutical diluents,excipients, or carriers (collectively referred to herein as "carrier"materials) suitably selected with respect to the intended form ofadministration, that is, oral tablets, capsules, softgels, elixirs,syrups, drops, and the like, and consistent with conventionalpharmaceutical practices.

For example, for oral administration in the form of tablets or capsules,a therapeutically effective amount of one or more compounds of thepresent invention can be combined with any oral non-toxicpharmaceutically acceptable inert carrier such as lactose, starch,sucrose, cellulose, magnesium stearate, dicalcium phosphate, calciumsulfate, mannitol, and the like, or various combinations thereof. Fororal administration in liquid forms, such as in softgels, elixirs,syrups, drops and the like, a therapeutically effective amount of theactive drug components can be combined with any oral non-toxicpharmaceutically acceptable inert carrier such as water, saline,ethanol, polyethylene glycol, propylene glycol, corn oil, cottonseedoil, peanut oil, sesame oil, benzyl alcohol, various buffers, and thelike, or various combinations thereof. Moreover, when desired ornecessary, suitable binders, lubricants, disintegrating agents, andcoloring agents ca also be incorporated in the mixture. Suitable bindersinclude starch, gelatin, natural sugars, corn sweeteners, natural andsynthetic gums such as acacia, sodium alginate, carboxymethylcellulose,polyethylene glycol, and waxes, or combinations thereof. Lubricants foruse in these dosage forms include boric acid, sodium benzoate, sodiumacetate, sodium chloride, and the like, or combinations thereof.Disintegrators include, without limitation, starch, methylcellulose,agar, bentonite, guar gum, and the like, or combinations thereof.Sweetening and flavoring agents and preservatives can also be includedwhere appropriate.

For intravascular, intraperitoneal, subcutaneous, or intramuscularadministration, one or more compounds of the present invention can becombined with a suitable carrier such as water, saline, agueousdextrose, and the like. For topical administration, such as forpsoriasis, therapeutically effective amounts of one or more compounds ofthe present invention can be combined with pharmaceutically acceptablecreams, oils, waxes, gels and the like. Regardless of the route ofadministration selected, the compounds of the present invention areformulated into pharmaceutically acceptable dosage forms by conventionalmethods known to those skilled in the art. The compounds can also beformulated using pharmacologically acceptable base addition salts.Moreover, the compounds or their salts may be used in a suitablehydrated form.

Regardless of the route of administration selected, a non-toxic buttherapeutically effective quantity of one or more compounds of thisinvention is employed in any treatment. The dosage regimen forpreventing or treating inflammatory conditions with the compounds ofthis invention is selected in accordance with a variety of factors,including the type, age, weight, sex, and medical condition of thepatient, the severity of the inflammatory condition, the route ofadministration, and the particular compound employed in the treatment. Aphysician or veterinarian of ordinary skill can readily determine andprescribe the effective amount of the drug required to prevent or arrestthe progress of the condition. In so proceeding, the physician orveterinarian could employ relatively low doses a first and subsequentlyincrease the dose until a maximum response is obtained. Daily dosages ofthe compounds of the invention are ordinarily in the range of about 1.0mg/kg up to about 30.0 mg/kg, (preferably in the range of about 2.0 to14.0 mg/kg (orally)).

The following examples illustrate the methods used to prepare thecompounds of this invention. These examples are given by way ofillustration only and are not meant to be construed as limiting theinvention in spirit or in scope, as many modifications in materials andmethods will be apparent from this disclosure to those skilled in theart.

In the following examples, and throughout this application, a wavy line( ) defines a substituent as an asymmetric carbon having R or Sstereochemistry or cis/trans isomers of a carbon carbon double bond. Inthe structures herein a bond drawn across a bond in a ring indicatesthat the bond can be to any available carbon atom of the ring structure.A series of dashes for a bond used in the structures herein indicatesthat such a bond may or may not be present. ##STR7##

EXAMPLE 1 ##STR8## The above acid chloride was prepared from terphthalicacid by reacting 0.5 g (3 mmoles) of terphthalic acid with 2 cc of[COCl]₂ (23.6 mmoles) in 10 cc of benzene and with one drop ofdimethylformamide. The reagents were mixed and warmed to 60° C. fortwenty four hours. The reaction mixture was cooled to room temperatureand the volatile components were removed in vacuo to give the abovecompound as a pale yellow solid. EXAMPLE 2

    CH.sub.3 (CH.sub.2).sub.11 C.tbd.C--TMS

The above compound was prepared by reacting an acetylene of the formulaCH₃ (CH₂)₁₁ C.tbd.CH (2.5 g. 12.87 mmoles) which was added to 25 cc oftetrahydrofuran (THF) and 50 mg of triphenylmethane (Ph₃ CH) which wasadded as an indicator. The solution was cooled to -30° C. and 1.6 molarn-butyllithium (n-BuLi) was added dropwise until the solution turnedred. Approximately 8.5 cc of n-BuLi was added. The solution was backtitrated with the acetylene compound until it became colorless. Thesolution was cooled to -78° C. and 2 cc (15.75 mmoles) of trimethylsilylchloride (TMS-Cl) was added. The solution was slowly warmed over aperiod of five hours to room temperature. The reaction was quenched withwater and extracted with hexane. The hexane was washed once with waterand once with brine and dried over magnesium sulfate (MgSO₄). Thetrimethylsilyl compound was isolated in an amount of 4.31 g (16.2mmoles).

EXAMPLE 3 ##STR9##

The above compound was prepared by reacting 3 mmoles of the acidchloride product from Example 1 with 0.8 g (3 mmoles) of the TMSacetylene product from Example 2. The acid chloride and theTMS-acetylene product were dissolved in 10 cc of dichloromethane andcooled to 0° C. To the reaction mixture was added 0.8 g (6 mmoles) ofaluminum chloride (AlCl₃) in small portions over ten minutes. Thereaction mixture was stirred for about 1.5 hours at 0° C. The reactionwas quenched with ice and the mixture was extracted three times withdiethyl ether. The extracts were combined and washed once with water andonce with brine (saturated sodium bicarbonate solution) and dried overmagnesium sulfate to yield 0.29 g of the above product.

HRMS (M⁺) calculated 342.2195; found 342.2196.

EXAMPLE 4 ##STR10## The above compound was prepared by mixing 3 g ofm-iodobenzoic acid (12.1 mmoles) and 3.12 g (15.0 mmoles) of anacetylene derivative of the formula ##STR11## with 0.085 g (0.121mmoles) of a palladium catalyst, Pd(PPh₃)₂ Cl₂ in 30 cc of Et₃ N. Thereaction vessel was purged with carbon monoxide. The reaction mixturewas heated under a carbon monoxide atmosphere (atmospheric pressure,balloon) in an oil bath at 80° C. for two hours. The reaction mixturewas cooled to room temperature. The volatile components were removed invacuo and the residue was taken up in 5% hydrochloric acid and extractedwith diethyl ether. The diethyl ether was washed once with 10%hydrochloric acid, twice with water, and once with a brine solution anddried over magnesium sulfate. The solvent was removed yielding 4.58 gmof the product. Chromatography on silica gel afforded 3.2 g of yellowsolid which was triturated with cold hexane to give a tan solid of 2.97g.

MP found: 77.5°-80.5° C.

Analysis calculated: 77.49; H, 9.05. Found: C, 77.22; H, 9.15.

EXAMPLE 5 ##STR12## The above compound was prepared by mixing one gram(4.03 mmoles) of m-iodobenzoic acid and 0.83 g (5 mmoles) of anacetylene derivative of the formula ##STR13## along with 0.028 g (0.04mmoles) of the palladium catalyst used in Example 4. The reagents weremixed in 10 cc of triethylamine. The reaction vessel was purged withcarbon monoxide. The reaction was performed under a carbon monoxideatmosphere (atmospheric pressure, balloon). The reaction mixture washeated in an oil bath at 80° C. for two hours. The reaction mixture wascooled to room temperature. The volatile components were removed invacuo and the residue was taken up in 5% hydrochloric acid (HCl) andextracted once with diethyl ether. The diethyl ether extract was washedonce with 10% HCl, twice with water and once with brine (NaCl) and driedover magnesium sulfate. The solvent was removed yielding a red yellowgum. After purification by silica gel chromatography 0.51 g of theproduct having the above structure was produced.

Analysis calculated: C, 76.40; H, 8.33. Found: C, 76.08; H, 8.42.

MP Found: 69°-71° C.

EXAMPLE 6 ##STR14##

A solution of 1 g (4.8 mmoles) of an acetylene having the formula

    H--C.tbd.C--(CH.sub.2).sub.12 CH.sub.3

in 25 cc of THF which contained a trace (20 mgs) of triphenylmethane wastreated with n-BuLi until the red color of the triphenylmethyl carbanionpersisted. A few drops of the acetylene was added to discharge the redcolor. The reaction was performed at 0° C. The reaction mixture wasstirred at 0° C. for 11/2 hours. The solution was added dropwise viasyringe to a cooled (0° C.) solution of phthalic anhydride (3.7 g, 25mmoles) in 50 ml of THF. The reaction mixture was stirred at roomtemperature for 21/2 days. The reaction mixture was quenched with 10%agueous HCl and extracted with ethyl acetate. The ethyl acetate extractwas washed once with water, once with brine and dried over MgSO₄. Afterremoval of the solvent, a gum was obtained. After purification by silicagel chromatography 0.104 g of the above product was recovered.

HRMS (M⁺): Calculated: 356.2352; Found: 356.2361.

EXAMPLE 7 ##STR15## A solution of 10 g (52.4 mmoles) of a bromo furanoicacid of the formula ##STR16## in 100 cc of methanol was prepared andcooled to 0° C. using an ice bath. 20 ml of concentrated sulfuric acidwas added dropwise over 15 minutes. The ice bath was removed and thereaction mixture stirred for eighteen hours at room temperatureovernight. The reaction mixture was poured onto 500 ml of water andextracted twice with diethyl ether. The extracts were combined andwashed once with water, once with NaHCO₃, once again with water andbrine and dried over magnesium sulfate. The solvent was removed to yield6 77 g of a pale yellow solid. EXAMPLE 8 ##STR17## The above compoundwas prepared by reacting 2.02 g (9.7 mmoles) of the acetylene compoundof the structure ##STR18## in 15 ml of THF at 0° C. To the solution wasadded 6.5 ml (10.4 mmoles) of 1.6 molar n butylithium. The reactionmixture was stirred for 25 minutes at 0° C. To the cooled milky solutionwas added 3 ml (11.1 mmoles) of ClSnBu₃ in a dropwise manner. Thesolution cleared to a pale yellow. The reaction was allowed to warm toroom temperature. The reaction mixture was diluted with dichloromethaneand washed with water and brine. The reaction mixture was dried oversodium sulfate, filtered and stripped to yield 5.13 g of the aboveproduct as a pale yellow oil. EXAMPLE 9 ##STR19## The above compound wasprepared by reacting 0.21 g (1.01 mmoles) of the product from Example 7and 0.5 g (1.01 mmoles) of the acetylene product from Example 8 in 5 ccof toluene. The palladium catalyst of Example 4, 7 mg (0.01 mmoles) wasadded to the reaction mixture. The reaction mixture was degassed andpurged three times with carbon monoxide. The reaction mixture was heatedto 100° C. using an oil bath and stirred for eight hours at 100° C. Thetoluene solution wa treated with saturated agueous potassium fluoridesolution for 1/2 hour. The mixture was then poured into diethyl etherand extracted twice with water and dried over magnesium sulfate. Afterfiltration and removal of the solvent, a brown gummy solid was obtained.Following chromatography on silica gel the above product was obtained,0.08 g (0.22 mmoles).

HRMS (M⁺) Found: 360.2303. Calculated: 360.2301.

EXAMPLE 10 ##STR20## The above compound was prepared by forming asolution of 2.5 gms.(13.1 mmoles) of a bromo furanoic acid in 25 ml oftetrahydrofuran (THF) which was cooled to -78° C. To the solution wasadded a 1.6 molar solution of n-butyl lithium in hexane (17.5 ml, 28mmoles) which was stirred at -78° for one hour. Dimethylformaide (DMF)was added in an amount of 2.4 ml (30 mmoles). The solution was allowedto warm to room temperature. The reaction mixture was quenched withwater and acidified with 10% hydrochloric acid. The resultant reactionmixture was extracted twice with ethyl acetate, the combined extractswere washed twice with water and once with brine and subsequently driedover magnesium sulfate. An orange solid was obtained after removal ofthe solvent in vacuo. Following chromatography on silica gel 0.73 gms.of the compound of the above formula was obtained. EXAMPLE 11 ##STR21##The above compound was prepared by forming a solution of 1.2 gms. (6mmoles) of an acetylene of the formula H.tbd.(CH₂)₁₂ CH₃ in 25 ml. THFwhich contained a catalytic amount (20 mg.) of triphenylmethane. Thesolution was cooled to -50° C. and then treated with 3.75 ml. (6 mmoles)of n-BuLi until the red color of the triphenylmethane anion persisted. Afew drops of the acetylene compound was added until the colordisappeared. An amount of 0.42 gms. of the product formed in Example 10in 10 ml. THF was added drop wise to the solution. The mixture waswarmed to 0° C. over one half hour. The mixture was quenched with waterand acidified with 10% hydrochloric acid. The agueous phase wasextracted twice with ethyl acetate. The extracts were combined andwashed twice with water and once with brine and were dried overmagnesium sulfate. Chromatography on silica gel yielded 0.80 gms. of apale yellow solid.

HRMS (M⁺) Calculated: 348.2301; Found: 348.2291.

EXAMPLE 12 ##STR22## The compound was prepared by reacting 0.145 g. (4.2mmoles of the product from example 11 in acetone (25 ml) and adding 1.5g. of activated MnO₂ portionwise over five minutes. The reaction mixturewas stirred for 24 hours at room temperature. The reaction mixture waspoured into 10% hydrochloric acid and extracted with ethyl acetate. Theextract was washed once with water and dried over magnesium sulfate.After the solvent was removed in vacuo a white solid remained. Followingchromatography on silica gel, 60 mg. of a white solid was recovered.

Analysis (for hydrate with 0.35 H₂ O); Calculated: C, 71.50; H, 8.77;Found: C, 71.55; H, 8.63.

EXAMPLE 13 ##STR23## The above compound was prepared by forming asolution of 0.83 gms. (4 mmoles) of an acetylene of the formulaH--C.tbd.C--(CH₂)₁₂ CH₃ in 25 ml. THF containing a catalytic amount (20mgs.) of triphenyl methane. The solution was cooled to -50° C., thentreated with 2.5 ml. of a 1.6 molar solution (4 mmoles) of n-BuLi inhexane until the red color of the triphenylmethane anion persisted. Afew drops of the acetylene was added until the color disappeared. Theresultant lithium acetylide preparation was cooled to -78° C. A solution(6 mmoles) of a diacid chloride of the formula ##STR24## was cooled to-78° C. and the -78° C. solution of lithium acetylide was added dropwisevia a cannula. The reaction was stirred for 10 minutes and quenched withwater and warmed to room temperature. The reaction mixture was pouredinto water and acidified with acetic acid (HOAc). The agueous solutionwas extracted twice with ethyl acetate and the extracts were washedtwice with water, once with brine and dried over magnesium sulfate.Following chromatography on silica gel 0.95 gms. of a product of theabove formula was recovered.

Analysis calculated: C, 73.92; H, 8.74; N, 3.92. Found: C, 73.64; H,8.79; N, 3.86.

EXAMPLE 14 ##STR25## The compound of the above formula was prepared inthe following manner. An acetylene solution was prepared by dissolving0.83 gms. 4 mmoles) of an acetylene of the formula H--C.tbd.C--(CH₂)₁₂CH₃ in 25 ml. of THF and 20 mgs. of triphenylmethane. The solution wascooled to -50° C. and treated with n BuLi until the red color of thetriphenylmethane anion persisted. A few drops of the acetylene was addeduntil the color disappeared. An aldehyde of the following formula##STR26## in an amount of 0.36 gms. (2 mmoles) was added dropwise to thesolution and the mixture was warmed to 0° C. The reaction mixture wasquenched with water and acidified with 10 % hydrochloric acid. Theagueous phase was extracted twice with ethyl acetate and the extractswere washed twice with water, once with brine and dried over magnesiumsulfate. Upon chromatography over silica gel 0.55 gms. (1.42 mmoles) ofa product of the above formula was recovered.

Analysis calculated: C, 74.19; H, 9.34; Found: C, 74.21; H, 9.47.

EXAMPLE 15 ##STR27## The above three compounds were prepared in thefollowing manner. An acid chloride of the structure ##STR28## wasprepared in a manner as recited in Example 1 in an amount of 15.05mmoles was dissolved in 50 ml. of dichloromethane and cooled to 0° C. Tothe solution was added 3.2 gms. (15 mmoles) of a acetylene of theformula H--C.tbd.C--(CH₂)₁₂ CH₃. 2 gms. of aluminum chloride was addedportionwise over one-half hour and the reaction mixture was stirred at0° C. for one hour. The reaction mixture was quenched with ice, dilutedwith ether and washed twice with water and dried over magnesium sulfate.Following separation by chromatography on silica gel 42 mg. and 30 mg ofthe isomers of the above formulae 15a and 15b were obtained and 23 mg.of the isomer of the above formula 15c wa obtained.

For 15a HRMS (M⁺) Calculated: 392.2118. Found: 392.2122.

For 15b HRMS (M⁺) Calculated: 392.2118. Found: 392.2121.

For 15c HRMS (M⁺) Calculated: 392 2118. Found: 392.2113.

EXAMPLE 16 ##STR29## A compound of the above structure was prepared byforming a solution of an acetylene of the formula H--C.tbd.C--(CH₂)₁₂CH₃ in 25 ml. of THF and 20 mgs. of triphenylmethane. The solution wascooled to -50° C. and treated with n-BuLi until the red color of thetriphenylmethane anion persisted. A few drops of acetylene was addeduntil the color disappeared. An aldehyde of the formula ##STR30## in anamount of 0.33 gms. (2 mmoles) in 5 ml. of THF was added dropwise to thesolution and the mixture was warmed to 0° C. The reaction mixture wasquenched with water and acidified with 10% hydrochloric acid. Theaqueous phase was extracted twice with ethyl acetate and the extractswere washed twice with water, once with brine and dried over magnesiumsulfate. Upon separation by chromatography on silica gel, 0.447 gms.(1.2 mmoles) of the product of the above formula was recovered as awhite solid.

Analysis calculated: C, 73.76; H, 9.15. Found: C, 73.54; H, 9.26.

EXAMPLE 17 ##STR31## The compound of the above formula was prepared inthe following manner. 0.1 gms. (0.27 mmoles) of the compound prepared inExample 16 was dissolved in 10 cc. of acetone. Activated manganesedioxide (1 gm.) was added portionwise. The reaction mixture was stirredfor 24 hours. The crude reaction mixture was filtered through celite andthe solvent evaporated. Following chromatography over silica gel 0.55gms. of a product of the above formula was produced as a white solid.

HRMS (M⁺) Calculated: 372 2300; Found: 372.2291.

EXAMPLE 18 ##STR32## The above compound was prepared in the followingmanner. Decadiyne in the amount of 6.71 gms. (15 mmoles) was dissolvedin 200 ml. of THF, along with 50 mgs. of triphenylmethane. The mixturewas cooled to -50° C. and 31.3 ml. (50 mmoles) of 1.6 molar n-BuLi inhexane was added until the red color of the triphenylmethane anionpersisted. Additional decadiyne was added until the red colordisappeared. The reaction mixture was warmed to -20° C. and stirred forone half hour. The reaction mixture was cooled to -40° C. and 9.9 gms.(50 mmoles) of iodohexane was added dropwise. Following the addition ofthe iodohexane, 50 ml. of HMPA was added dropwise and the reactionmixture was stirred and allowed to warm to room temperature. Thereaction mixture was quenched with water and poured into hexane andwashed with four washings of water, one with brine and dried overmagnesium sulfate. The crude product contained the diyne of the formulaH--C.tbd.C--(CH.sub. 2)₆ --C.tbd.C--C₅ H₁₁.

A lithium acetylide was prepared by treating the crude diyne with 14.7mmoles of n-BuLi at -20° C. until the color of the triphenylmethaneanion was present. Triphenylmethane (25 mgs) was added. An excess of theacetylene was added until the color disappeared. The reaction mixturewas cooled to -50° C. and 1 gm. (6.7 mmoles) of an aldehydic acid of theformula ##STR33## in 10 mls. of THF was added dropwise. The reactionmixture was warmed to room temperature over 1 hour and the reaction wasquenched with water. The bulk of THF was removed in vacuo and theresidue partitioned between diethyl ether and water The diethyl etherlayer was dried over magnesium sulfate. Upon chromatography 1.11 gms.(3.13 mmoles of the product of the above formula was produced as ayellow oil which solidified on standing.

Analysis calculated: C, 77.93; H, 8.53. Found: C, 78.02; H, 8.43.

HRMS (M⁺) Calculated: 354.2195. Found: 354.2195.

EXAMPLE 19 ##STR34## The compound of the above formula was prepared inthe following manner. A solution was prepared by dissolving 0.125 gms.(0.35 mmoles of the product from Example 18 in 15 ml. of acetone. To thesolution was added 1 gm. of manganese dioxide (MnO₂). The manganesedioxide was added portionwise over a period of 15 minutes. The reactionmixture was stirred for 24 hours at room temperature. The reactionmixture was poured into 10% hydrochloric acid and extracted with ethylacetate. The extract was washed once with water and dried over magnesiumsulfate. The solvent was removed in vacuo and the product was isolatedusing chromatography on silica gel to provide 0.90 gms. of a product ofthe above formula.

Analysis calculated: C, 78.38; H, 8.01. Found: C, 78.53; H, 8.18.

EXAMPLE 20 ##STR35## A compound of the above formula was prepared in thefollowing manner. 10 mgs. (0.028 mmoles) of the compound prepared inExample 19 was mixed with 2.0 ml. of hexane and 1 mg. of a Lindlarcatalyst and 0.01 ml. of a quinoline. The reagents were mixed and thereaction vessel was degassed 3 times with hydrogen delivered from aballoon. The reaction mixture was monitored closely via TLC and stirredat room temperature. The reaction vessel was evacuated and purged threetimes with argon. The reaction mixture was then filtered through celite.The filtrate was washed once with 10% hydrochloric acid, once withwater, once with saturated sodium bicarbonate, once with saturatedsodium chloride, and dried over magnesium sulfate. Following separationby chromatography 0.008 gms. of a product of the above formula wasrecovered as a mixture of E and Z isomers about the enone double bond.

HRMS (MH⁺) Calculated: 357.2430. Found: 357.2432.

EXAMPLE 21 ##STR36## A compound of the above structure was prepared inthe following manner. To a solution of 0.1 gms. of the product fromExample 18 (0.282 mmoles) in 1 ml. of DMF was added 0.1 gms. (1.5mmoles) of imidazole, followed by 0.19 gms. (0.7 mmoles) oft-butyldiphenylsilyl chloride (TBDPS-Cl) at room temperature underargon. The solution was stirred for 4 hours at room temperature. Thereaction mixture was diluted with 200 ml. of diethyl ether and washed 3times with water (30 ml. each), once with brine and dried over magnesiumsulfate. Removal of the solvent yielded an oil which upon separation bychromatography on silica gel yielded 0.11 gms. of a product of theformula ##STR37## This product, 0.11 g (0.13 mmoles) was mixed with 10ml. of hexane, 5 mgs. of a Lindlar catalyst and 0.05 ml of quinoline.The reaction mixture was stirred at room temperature under hydrogen andafter 1 hour TLC indicated the reaction was complete. The reactionvessel was evacuated and purged 3 times with argon. The reaction mixturewas filtered through celite, diluted with hexane and washed once with 5%hydrochloric acid, once with water, once with brine and dried overmagnesium sulfate. The resultant product produced was 0.11 gms of aproduct of the formula ##STR38## A solution of this product was preparedby dissolving 0.11 gms. in 10 ml. of THF, which was then treated with1.5 ml. of 1 molar n-Bu₄ NF in THF. After the starting material wasconsumed (via TLC), the reaction mixture was diluted with diethyl etherand washed with water and dried over magnesium sulfate. Followingchromatography 0.037 gms. (0.103 mmoles) of the above identified productwas recovered.

HRMS (MH⁺) Calculated: 357.2430. Found: 357.2444.

EXAMPLE 22 ##STR39## A compound of the above formula was prepared in thefollowing manner. The compound prepared in Example 21, 0.030 gms., wasdissolved in 3 ml. of acetone and 0.3 gms. of MnO₂ was added portionwiseover 5 minutes. The reaction mixture was stirred for 24 hours at roomtemperature. The reaction mixture was then poured into 10% hydrochloricacid and extracted with ethyl acetate. The extract was washed once withwater and dried over magnesium sulfate. After the solvent was removed invacuo the resultant product was separated by chromatography over silicagel, yielding 0.021 gms. of product of the above formula.

HRMS (M⁺) Calculated 354.2192. Found 354.2195.

EXAMPLE 23 ##STR40## A compound of the above formula was prepared in thefollowing manner. To a cooled (0° C.) solution of 5.5 ml. (39.2 mmoles)of diisopropylamine in 50 ml. of THF was added 20.5 ml. of 1.6 molarBuLi (32.8 mmoles) to make 32.8 mmoles of lithium diisopropylamide(LDA). The reaction mixture was stirred for one-half hour at 0° C. andcooled to -78° C. To the mixture was added 2.1 gm (16.4 mmoles) of2-thiophene carboxylic acid in 25 ml. THF. Additional THF was added toincrease the volume to 200 ml. and the reaction mixture was stirred forone-half hour. DMF was added in an amount of 1.3 ml. (16.8 mmoles). Thereaction mixture was warmed to room temperature and stirred for 11/2hours. The reaction mixture was quenched with water and acidified with1N hydrochloric acid and extracted with ethyl acetate. The organicextracts were combined and dried over magnesium sulfate. The resultantmixture was filtered and stripped to yield a yellow solid. Separationusing chromatography on silica eluting with ethyl acetate/hexane/1%acetic acid provided 1.3 gms. of a yellow solid of the above formula. MP160°-163°.

Analysis calculated: C, 46.15; H, 2.58. Found: C, 46.11; H, 2.82.

EXAMPLE 24 ##STR41## The compound with the above structure was preparedin the following manner. An acetylene of the formula H--C.tbd.C--(CH₂)₁₂CH₃ in an amount of 549.3 mg. (2.6 mmoles) in 15 ml. of THF was cooledto -20° . To the solution was added 1.6 ml. (2.6 mmoles) of n-BuLi. Thereaction was stirred for one half hour and 203.4 mg. (1.3 mmoles) of theproduct from Example 23 in 10 ml. of THF was added. The mixture wasstirred and maintained at -20° for 15 minutes and allowed to warm to 0°C. and stirred for one-half hour. The reaction mixture was quenched withwater and acidified with 10% hydrochloric acid and extracted with ethylacetate. The combined organic extracts were dried over magnesiumsulfate. The extract was filtered and stripped to yield a yellow oilwhich solidified upon standing. The solid was dissolved in ethyl acetateand filtered through silica gel eluting with 100% hexane followed byethyl acetate in 1% acetic acid. The ethyl acetate fraction yielded392.8 mg. of the above compound as a yellow solid having a melting pointof 75°-90°.

Analysis calculated: C, 69.19; H, 8.85. Found: C, 69.18; H, 9.02.

EXAMPLE 25 ##STR42## A compound of the above structure was formed in thefollowing manner. 7.2 mg (0.020 mmoles) of the compound prepared inExample 24 was dissolved in 1 ml. acetone. To the solution was added 72mg. (0.83 mmoles) of activated manganese dioxide. The reaction mixturewas stirred vigorously at room temperature overnight. The reactionmixture was poured into 10% hydrochloric acid. The aqueous phase wasextracted with ethyl acetate. The organic washes were combined and driedover magnesium sulfate. The organic phase was filtered and stripped toyield 6 mg. (0.010 mmoles) of the above product as a white solid.

HRMS (M⁺) Calculated: 362.1916. Found: 362.1910.

EXAMPLE 26 ##STR43## A compound of the above formula was prepared in thefollowing manner. m-Iodobenzoic acid was reacted in an amount of 569mgs. 2.29 mmoles) with 440 mg. (2.30 mmoles) of1-(3-dimethylaminopropyl)-3-ethyl carbodiimide.HCl in 25 ml. of DMF. Tothe reaction mixture was added 0.35 ml. (2.51 mmoles) of Et₃ N and 361mg. (2.30 mmoles) of NH₂ SO₂ (C₆ H₅). The reaction mixture was stirredat room temperature for 12 hours. The reaction mixture was poured intowater and extracted with ethyl acetate. The organic extracts werecombined and washed with brine and dried over magnesium sulfate.Following chromatography on silica gel eluting with ethyl acetate/hexane/1% acetic acid, 126 mg. of a white solid of the product with theabove formula was recovered. EXAMPLE 27 ##STR44## A compound with theabove formula was prepared in the following manner. The compound fromExample 26 in an amount of 77 mg. 0.20 mmoles) was combined with 49 mg.(0.24 mmoles) of an acetylene of the formula H--C.tbd.C--(CH₂)₁₂ CH₃ and3 mg. (0.004 mmoles) of Pd(Ph₃)₂ Cl₂ in 2 ml. of Et₃ N. The mixture washeated in an oil bath at about 80° C. The reaction vessel was purged 4times with carbon monoxide from a carbon monoxide balloon and permittedto react with stirring under carbon monoxide for 2.5 hours. The reactionmixture was then stirred at room temperature for about 60 hours. Thesolvent was evaporated under nitrogen. The residue was dissolved inethyl acetate and washed with successive washes of 10% hydrochloricacid, water and brine. The organic layer was dried over magnesiumsulfate, filtered and stripped to yield a brown oil. Upon chromatographyon silica gel, 35 mgs. of a compound with the above formula wasrecovered as a brown oil.

HRMS (MNH₄ ⁺) Calculated: 513.2787. Found: 513.2761.

EXAMPLE 28 ##STR45## A compound having the above formula was prepared inthe following manner. A solution of m-cyanobenzaldehyde was prepared bydissolving 2.0 gms. 15.2 mmoles) of the benzaldehyde along with 2.98gms. 45.8 mmoles) of NaN₃ and 2.9 gms. (21.1 mmoles) of Et₃ N.HCl, allof which were dissolved in 50 ml. of 1-methyl-2-pyrrolidinone. Thereaction mixture was refluxed under argon. After 1 hour and 45 minutesthe reaction mixture was cooled to room temperature and poured into 200ml. of water and acidified with 10% hydrochloric acid. The reactionmixture was extracted with successive ethyl acetate washes. The ethylacetate extracts were combined and washed with brine and dried overmagnesium sulfate. The ethyl acetate extract was chromatographed throughsilica gel, yielding 0.3 gms. of the product having the above formula asa white solid. EXAMPLE 29 ##STR46## A compound of the above formula wasprepared in the following manner. A solution of 5.03 g (24.1 mmoles) ofan acetylene of the formula HC.tbd.C(CH₂)₁₂ CH₃ was dissolved in 100 ml.THF and cooled to -30° C. To the solution was added 15 ml. (24 mmoles)of a 1.6 molar n-BuLi solution which was added dropwise. The reactionmixture was stirred for 15 minutes at which time 2.1 gms. (12.0 mmoles)of the product from Example 28 dissolved in 75 ml. of THF was addeddropwise. The solution was stirred and maintained at -30° C. forone-half hour, then warmed to room temperature. The reaction mixture wasquenched with water and acidified with 10% hydrochloric acid. The layerswere separated and the organic phase was washed with brine and driedover sodium sulfate. The layer was filtered and stripped to yield ayellow solid which upon chromatography over silica gel yielded 3.75 gms.of the above product as a white solid.

Analysis calculated: C, 72.21; H, 8.96; N, 14.65. Found: C, 72.03; H,9.00; N, 14.77.

EXAMPLE 30 ##STR47## The compound having the above formula was preparedin the following manner. A solution was prepared by dissolving 36.2 mgs.(0.095 mmoles) of the product from Example 29 in acetone. To thesolution was added 360 mgs. (4.14 mmoles) of activated manganesedioxide. The reaction mixture was stirred vigorously at room temperatureovernight. The reaction mixture was poured into 10% hydrochloric acidThe aqueous phase was extracted with ethyl acetate and the organicwashes were combined and dried over magnesium sulfate. The organic phasewas filtered and stripped to yield 18 mgs. of a pale yellow solid of acompound having the above formula.

Analysis calculated: C, 72.59; H, 8.48; N, 14.72. Found: C, 72.19; H,8.66; N, 14.13.

ASSAY FOR LTB₄ AND PGE₂ PRODUCTION BY HL-60 CELLS

HL-60 cells were induced to differentiate into granulocytes by a 4 dayincubation with 0.8% (v/v) N,N-dimethyl formamide as disclosed inFontana et al., Proc. Natl. Acad. Sci. 78 (6):3863-3866 (1981); Agins etal., Biochem. Biophys. Res. Comm. 126, 143-149 (1985); and Bonser etal., Biochemistry 20: 5297-5301 (1981). Prior to performing the assay,differentiated HL-60 cells were washed once with Hanks' balanced saltsolution containing 0.35 mg/ml sodium bicarbonate and 10 mM HEPES pH7.35 (HBSS). HL-60 cells (3×10⁶ cells/ml) were pre-incubated with thecompound tested or a control vehicle at 37° C. for 10 minutes, followedby 5 minute incubation with 5×10⁶ M calcium ionophore A23187 in a finalvolume of 1.0 ml. After incubation, the cells were pelleted bycentrifugation and the LTB₄ and PGE₂ in the supernatent were quantifiedby radioimmunoassay. IC₅₀ values (means + /- S.E.) for compounds hereinthat were tested are shown in the following Table and represent theconcentrations of the compound required to inhibit 50% of LTB₄ or PGE₂production by HL-60 cells stimulated with the calcium ionophore A23187.

HUMAN SYNOVIAL FLUID PHOSPHOLIPASE A₂ (HSF-PLA₂) ASSAY

Human synovial fluid phospholipase A₂ was purified approximately 5000fold following the procedures of Franson et al., Lung 160, 275-284(1982) and Fawzy et al., Bio Phys. J. 49, 533a (1986). Followingpurification the enzyme activity was measured by established methodologyusing [¹⁴ C]-oleate-labeled, autoclaved E. coli as the substrate as alsoshown in the above noted references. The assay was performed in a finalvolume of 100 ml containing 50 mM HEPES (pH 7.0), 150 mM NaCl, 5 mMCaCl₂, 7 mM [¹⁴ C]-oleate-labeled E. coli phospholipid and with orwithout the compound from one of the examples herein undergoing anassay. The compound or control vehicle was pre-incubated with the PLA₂for 5 minutes followed by addition of the E. coli substrate to initiatethe reaction. The reaction was maintained at 37° C. for 30 minutes andthen terminated by the addition of 2 ml tetrahydrofuran (THF). Thereaction product, [¹⁴ C]-oleic acid, was extracted using a 1 ml BondElut-NH₂ Solid phase extraction column. The IC₅₀ value for the compound(mean +/- S.E.) is given in the following Table and represents theconcentration of the compound required to inhibit 50% of the PLA₂activity.

                  TABLE                                                           ______________________________________                                                                 LTB4 Biosynthesis                                                             inhibition (HL60                                     Example # HSF-PLA2 IC50 uM                                                                             cells) IC50 uM                                       ______________________________________                                         3        14             0.2                                                   4        11             0.8                                                   6        13             --                                                   11        28             2.4                                                  12        34             0.5                                                  13          2.5          0.8                                                  16        11             8                                                    17        33             0.4                                                  18        28             --                                                   24        18             0.9                                                  25        15             0.4                                                  27        14             --                                                   29        42             --                                                   30        11             0.9                                                  ______________________________________                                    

What is claimed is:
 1. A compound of the formula: ##STR48## or apharmaceutically acceptable salt thereof wherein X is oxygen;wherein Ywhen present is hydrogen or halogen; wherein R¹ is alkyl of 1 to 20carbon atoms, alkenyl of 2 to 20 carbon atoms, or alkynyl of 2 to 20carbon atoms; wherein R is --CO₂ R², tetrazole, methylsulfonamide orbenzenesulfonamide; wherein R² is hydrogen, alkyl of 1 to 6 carbons or apharmaceutically acceptable cation; and wherein R³ is hydroxyl orhalogen.
 2. A compound according to claim 1 of the formula: ##STR49## ora pharmaceutically acceptable salt thereof.
 3. A compound according toclaim 2 wherein R¹ is an alkyl from 1 to 20 carbon atoms.
 4. A compoundaccording to claim 2 of the formula: ##STR50##
 5. A compound accordingto claim 2 of the formula: ##STR51##
 6. A compound according to claim 2of the formula: ##STR52##
 7. A pharmaceutical composition comprising atherapeutically of prophylactically effective amount of a compoundaccording to claim 1 in a pharmaceutically acceptable carrier.
 8. Apharmaceutical composition according to claim 7 wherein said compositionis in oral dosage form.
 9. A method of treating inflammatory conditionsin mammals comprising administering to a patient in need of suchtreatment, a therapeutically effective amount of a compound according toclaim 7 in a pharmaceutically acceptable carrier.
 10. A method ofpreventing an inflammatory attach in mammals comprising administering toa patient susceptible to such attach a prophylactically effective amountof a compound according to claim 1 in a pharmaceutically acceptablecarrier.